Survivin多表位真核表达载体的构建及其在树突状细胞中的表达  

Construction of the eukaryotic expression vector encoding multi-epitope fusion protein of human survivin and its expression in dendritic cells

作　　者：王效远[1] 宋文哲[1] 李向农[1] 郭萌[1] 梁骁[1] 李莹[1] 

机构地区：[1]徐州医学院附属医院普通外科,江苏徐州221002

出　　处：《细胞与分子免疫学杂志》2011年第5期498-500,共3页Chinese Journal of Cellular and Molecular Immunology

摘　　要：目的:构建人生存素(survivin)的多表位真核表达载体,并在人树突状细胞(DC)中进行表达。方法:人工合成含4个survivin的HLA-A2类限制性CD8+CTL表位和一个CD4+Th表位的重组cDNA片段,克隆到pBluescript II SK(+)载体上,测序正确后将目的基因连接到真核表达载体pIRESneo3.0中,构建的重组真核表达质粒pPIRESneo3.0-survivin(4)/Th。将其转染人DC并筛选,进行稳定表达。结果:构建了含有survivin的多表位真核表达载体,并成功地转染了人DC。结论:成功地构建了含有survivin的多表位真核表达载体,并在人DC中稳定表达,为进一步研究多表位肿瘤疫苗奠定基础。AIM： To construct a eukaryotic expression vector encoding the multi-epitope fusion protein of human survivin,and express it in human dendritic cells.METHODS： Recombinant cDNA sequence encoding four HLA-A2-restricted CD8＋ CTL epitopes and a CD4＋ Th epitope was synthesized and cloned into pBluescript II SK（＋） vector.After confirmed by sequencing,the cDNA fragment was inserted to eukaryotic expression vector pIRESneo3.0 to generate the recombinant plasmid pPIRESneo3.0-survivin（4）/Th.The pPIRESneo3.0-survivin（4）/Th was then transfected into human dendritic cells and the transfectants were selected for stable expression.RESULTS： The eukaryotic expression vector encoding the multi-epitope fusion protein of survivin was constructed,and successfully transfected into human dendritic cells.CONCLUSION： The eukaryotic expression vector encoding the multi-epitope fusion protein of survivin has been constructed successfully,and stably expressed in human dendritic cells,which provides clues for further research on multi-epitope cancer vaccine.

关 键 词：SURVIVIN 多表位 树突状细胞 肿瘤疫苗 

分 类 号：R392.11[医药卫生—免疫学]